1. Field of the Invention
The present invention relates to a method and an apparatus for observing three-dimensional localizations of in vivo expressed genes as well as a method and an apparatus for observing minute three-dimensional localizations of in vivo expressed genes, and more particularly to a method and an apparatus for observing three-dimensional localizations of in vivo expressed genes as well as a method and an apparatus for observing minute three-dimensional localizations of in vivo expressed genes used suitably in the case when an expressed situation in a variety of genes is observed in living organisms such as animals and plants.
2. Description of the Related Art
Heretofore, such a procedure that a living organism to be used for a specimen (an expression “living organism to be used for specimen” is hereinafter referred optionally to as “living specimen” in the present specification) is sliced, and the sliced living specimen is observed, whereby a two-dimensional configuration of expressed genes is observed in the living specimen has been adopted in case of observing a localization of genes expressed inside a living organism, i.e., in vivo expressed genes.
Furthermore, it becomes possible to discriminate expressed sites of specific genes inside a living specimen in accordance with such a manner that a marker (label), which can be detected at the time of gene expression, is incorporated in genes being objects to be observed, and then, a resulting living genetic recombinant specimen containing the genes into which the marker has been incorporated is observed in the case where the observation technique as described above is applied.
In this case, fluorescent materials, luminescent materials, coloring matters and the like may be used for such marker as described above. Specific examples thereof include GFP, EGFP, YFP, RFP, luciferin, melanin pigment and the like.
However, the observing techniques as described above involve such problems that a range, which can be observed, is limited in the case where an entire living specimen is looked through, because a stereoscopic microscope is used in comparatively broad macro observation as well as that observation is difficult upon a minute site, so that resolving power becomes poor.
For this reason, there has been such a problem that it is very difficult to observe localizations of in vivo expressed genes in an entire individual living organism according to a conventional observing technique.
On hand, while there is “in-situ Hybridization” as a method for observing sliced expressed genes, it is necessary for infiltrating a marker such as fluorescent materials, coloring matters and the like into the interior of a living specimen in this method, so that it is difficult to dye homogeneously the interior of the living specimen.
Besides, when the observation as described above is implemented, such an observing technique wherein a confocal laser microscope is used has been applied in case of micro observation by which a minute region is observed. In this case, however, a range, which can be observed by such technique as described above, is narrow, in other words, an extent of which is only up to around 100 μm from its surface of a living specimen to be observed in the thick direction thereof.
Therefore, there has been such a problem that it is extremely difficult to observe three-dimensional localizations of in vivo expressed genes in an entire individual living organism.